Encapsulated droplet interface bilayers as a platform for high-throughput membrane studies

Whilst it is highly desirable to produce artificial lipid bilayer arrays allowing for systematic high-content screening of membrane conditions, it remains a challenge due to the combined requirements of scaled membrane production, simple measurement access, and independent control over individual bilayer experimental conditions. Here, droplet bilayers encapsulated within a hydrogel shell are output individually into multi-well plates for simple, arrayed quantitative measurements. The afforded experimental throughput is used to conduct a 2D concentration screen characterising the synergistic pore-forming peptides Magainin2 and PGLa. Maximal enhanced activity is revealed at equimolar peptide concentrations via a membrane dye leakage assay, a finding consistent with models proposed from NMR data. The versatility of the platform is demonstrated by performing in situ electrophysiology, revealing low conductance pore activity (similar to 15 to 20 pA with 4.5 pA sub-states). In conclusion, this array platform addresses the aforementioned challenges and provides new and flexible opportunities for high-throughput membrane studies. Furthermore, the ability to engineer droplet networks within each construct paves the way for lab-in-a-capsule approaches accommodating multiple assays per construct and allowing for communicative reaction pathways.

Automated summary

In this study, a high-throughput membrane platform was developed using droplet bilayers encapsulated within a hydrogel shell. Through concentration screening and electrophysiology experiments, maximal activity enhancement at specific peptide concentrations and low conductance pore activity were revealed.